Celltiter 96 aqueous one mts assay

Manufactured by Promega

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The CellTiter 96 AQueous One MTS assay is a colorimetric method for determining the number of viable cells in proliferation or cytotoxicity assays. It measures the bioreduction of a tetrazolium compound into a colored formazan product, which can be quantified using a spectrophotometer.

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MTS assay (792 citations) CellTiter 96 (150 citations) CellTiter 96 AQueous (88 citations) MTS CellTiter 96 (2 citations)

6 protocols using celltiter 96 aqueous one mts assay

Plasmid-Mediated Prodrug Activation

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1,4-butanediol diacrylate (B4; CAS 1070-70-8), 3-amino-1-propanol (S3, CAS 156-87-6), 4-amino-1-butanol (S4, CAS 13325-10-05), 5-amino-1-pentanol (S5, CAS 2508-29-4), and 1-(3-aminopropyl)-4-methylpiperazine (E7, CAS 4572-031) were purchased from Alfa Aesar (Ward Hill, MA, USA); 1,5-pentanediol diacrylate (B5, CAS 36840-85-4) was purchased from Dajac Laboratories (Feasterville-Trevose, PA, USA); 2-(3-aminopropylamino)ethanol (E6; CAS 4461-39-6) was purchased from Sigma Aldrich (St. Louis, MO, USA). Lipofectamine™ 2000 and Lipofectamine 3000™ were purchased from ThermoFisher (Waltham, MA, USA). 25 kD branched poly(ethylenimine) was purchased from Sigma Aldrich (St. Louis, MO, USA). The pEGFP-N1 plasmid (GFP) was purchased from Elim Biopharmaceuticals (Hayward, CA, USA) and the herpes simplex virus type 1-derived thymidine kinase (HSVtk) gene was cloned into the pcDNA3.1 vector; both plasmids were amplified by Aldeveron (Fargo, ND, USA). For cell staining in in vitro cell killing assays, Hoechst 33342 and propidium iodide were purchased from Invitrogen (Carlsbad, CA, USA). CellTiter 96 AQ_ueous One MTS assay was purchased from Promega (Madison, Wisconsin, USA). Ganciclovir was purchased from Invivogen (Carlsbad, CA, USA).

Choi J., Rui Y., Kim J., Gorelick N., Wilson D.R., Kozielski K., Mangraviti A., Sankey E., Brem H., Tyler B., Green J.J, & Jackson E.M. (2019). Nonviral Polymeric Nanoparticles for Gene Therapy in Pediatric CNS Malignancies. Nanomedicine : nanotechnology, biology, and medicine, 23, 102115.

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Multifunctional Polymer Synthesis and Evaluation

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1,4-butanediol diacrylate (B4), 4-amino-1-pentanol (S4), 5-amino-1-pentanol (S5), 1-(3-aminopropyl)-4-methylpiperazine (E7)
(Alfa Aesar), 1,5-pentanediol diacrylate (B5) (Monomer Polymer & Dajac Labs), 2-methylpentane-1,5-diamine (E4) (TCI
America), 2-(3-aminopropylamino)ethanol (E6) (Fluka), poly(ethylene glycol) methyl ether thiol (800 Da), branched 25 kDa
poly(ethylenimine) (PEI) (Sigma-Aldrich), α-Mercaptoethyl-ω-methoxy polyoxyethylene (5000 Da) (NOF America
Corporation), and cell culture media components were purchased and used as received. pEGFP-N1 (EGFP) DNA (purchased from Elim
Biopharmaceuticals and amplified by Aldevron, Fargo, ND), ganciclovir (Invivogen, San Diego, CA), Label IT-Tracker Cy3 kit (Mirus
Bio LLC), and CellTiter 96 AQueous One MTS assay (Promega, Fitchburg, WI) were obtained from commercial vendors and used per
manufacturer’s instructions. HSV-tk gene was cloned into the pcDNA3.1 vector (Life Technologies) and amplified (Aldevron,
Fargo, ND).

Kim J., Kang Y., Tzeng S.Y, & Green J.J. (2016). Synthesis and application of poly(ethylene glycol)-co-poly(β-amino ester) copolymers for small cell lung cancer gene therapy. Acta biomaterialia, 41, 293-301.

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Polymer Synthesis and Characterization

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1,4-butanediol diacrylate (B4), octylamine (S8m), decylamine (S10m), pyrene (Sigma-Aldrich), 1,6-hexanediol diacrylate (B6), 1-(3-aminopropyl)-4-methylpiperazine (E7) (Alfa Aesar), dimethyl sulfoxide (DMSO), dimethyl formamide (DMF), hexane, methoxy poly(ethylene glycol) thiol (2 kDa and 800 Da) (Laysan Bio, Inc), and Verteporfin (VP) (U.S. Pharmacopeial Convention, Inc.) were purchased and used as received. CellTiter 96 AQueous One MTS assay (Promega, Fitchburg, WI) was used per manufacturer’s instructions.

Kim J., Shamul J.G., Shah S.R., Shin A., Lee B.J., Quinones-Hinojosa A, & Green J.J. (2018). Verteporfin-loaded poly(ethylene glycol)-poly(beta-amino ester)- poly(ethylene glycol) triblock micelles for cancer therapy. Biomacromolecules, 19(8), 3361-3370.

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Biomaterial Platform for Cell Engineering

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1,4-butanediol diacrylate (B4), 4-amino-1-butanol (S4), 5-amino-1-pentanol (S5), 1-(3-aminopropyl)-4-methylpiperazine (E7) (Alfa Aesar), pentane-1,3-diamine (E3) (TCI America), 2-(3-aminopropylamino)ethanol (E6) (Fluka), poly(ethylene glycol) methyl ether thiol (800 Da and 2000 Da), (Sigma-Aldrich), α-mercaptoethyl-ω-methoxy polyoxyethylene (5000 Da) (NOF America Corporation), and cell culture media components were purchased and used as received. HSV-tk gene cloned into the pcDNA3.1 vector (Life Technologies), and pEGFP-N1 (EGFP) and pDsRed (DsRed) DNA (Elim Biopharmaceuticals) were amplified by Aldevron. Ganciclovir (Invivogen), Label IT-Tracker Cy5 kit (Mirus Bio LLC), and CellTiter 96 AQueous One MTS assay (Promega) were obtained from commercial vendors and used per manufacturer’s instructions.

Kim J., Mondal S.K., Tzeng S.Y., Rui Y., Al-kharboosh R., Kozielski K.K., Bhargav A.G., Garcia C.A., Quiñones-Hinojosa A, & Green J.J. (2020). Poly(ethylene glycol)-poly(beta-amino ester)-based nanoparticles for suicide gene therapy enhance brain penetration and extend survival in a preclinical human glioblastoma orthotopic xenograft model. ACS biomaterials science & engineering, 6(5), 2943-2955.

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MTS Cytotoxicity Assay for RAW Cells

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Compound toxicity against host cells was determined by CellTiter 96
AQ_ueous One MTS Assay (Promega, Madison, WI). RAW cells were
harvested by gently scraping in PBS, resuspended in DMEM at a concentration of
1×10⁵ cells/ml, and seeded into 96-well tissue culture
plates (Corning) at 10⁴ per well. Cells were allowed to adhere to the
plate overnight. Compounds were added to a concentration of 10 μM and
serially diluted in steps of 1:3, keeping a constant volume of 100
μl/well. Vehicle addition (0.1% DMSO) and media only controls
were included in each plate. Cells were incubated for 48 h at 37°C,
5% CO₂ after which 20 μl of MTS reagent was added to
wells and plates were incubated for a further 4 h at 37°C, 5%
CO₂. The absorbance of the reaction product was read at 490 nm in
an EL-800 Plate Reader (Biotek). Three technical replicates were performed for
each sample and the assay was performed three separate times.

Simpson C., Jones N.G., Hull-Ryde E.A., Kireev D., Stashko M., Tang K., Janetka J., Wildman S.A., Zuercher W.J., Schapira M., Hui R., Janzen W, & Sibley L.D. (2015). Identification of small molecule inhibitors that block the Toxoplasma gondii rhoptry kinase ROP18. ACS infectious diseases, 2(3), 194-206.

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Synthesis and Transfection of BR6 Monomer

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All chemicals used for the synthesis of monomer BR6 were purchased from Sigma-Aldrich Chemical Co. (St. Louis, MO) and used without further purification. All other monomers were purchased from Alfa Aesar (Ward Hill, MA). Lipofectamine 2000 and Opti-MEM I were purchased from Invitrogen (Carlsbad, CA) and used according to manufacturer’s instructions. Ambion Silencer eGFP and Ambion Silencer Negative Control #1 siRNA were purchased from Life Technologies. CellTiter 96 AQ_ueous One MTS assay was purchased from Promega (Fitchburg, WI) and used according to manufacturer’s instructions.

Kozielski K.L., Tzeng S.Y., Hurtado De Mendoza B.A, & Green J.J. (2014). Bioreducible Cationic Polymer-Based Nanoparticles for Efficient and Environmentally Triggered Cytoplasmic siRNA Delivery to Primary Human Brain Cancer Cells. ACS Nano, 8(4), 3232-3241.

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